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KMID : 0578319920020020207
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Molecules and Cells
1992 Volume.2 No. 2 p.207 ~ p.211
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Expression of Mutated δ-Endotoxin Gene of Bacillus thuringiensis subsp. Tenebrionis in E. coli and Insecticidal Activity against Coleopteran Insects
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Rhim Seong-Lyul
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Abstract
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A cloned S-endotoxin gene from Bacillus thuringiensis subsp. tenebrionis (Btt) was mutated at 5´-end region by site directed mutagenesis. The mutation results in creation of a new BamHI restriction site. For general cloning and further researches such as analysis of gene expression, the promoter region was replaced with a synthesized oligonucleotide containing Smal, BgQI and BamHI restriction sites. In the synthesized sequence, a ATG-start codon was included before the new BamHI site. This sequence was subsquently fused to lacZ´-promoter. The expression of two proteins indicated a second ribosome binding site of the toxin encoding sequence. It was found by the Western blot analyses that the expression of intact and modified Btt-toxin genes showed no significant differences in E. coli. Furthermore, biotest with extract of E. coli transformant by mutated Btt-toxin gene showed toxin activity against coleopteran insect larvae.
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